胶原酶消化法培养大鼠远端肺静脉平滑肌细胞

目的：探索大鼠发病机制远端肺静脉平滑肌细胞（PVSMC）的原代培养方法,为体外研究肺血管疾病提供实验材料.方法：采用显微操作和Ⅰ型胶原酶消化法原代培养大鼠远端PVSMC,对培养的细胞进行形态学观察、平滑肌α-actin免疫荧光染色鉴定,并用激光扫描共聚焦显微镜计算纯度.结果：镜下培养的细胞呈典型的＂峰-谷＂状生长,平滑肌细胞特异的平滑肌α-actin蛋白阳性表达,细胞纯度达98.5%.结论：用胶原酶消化法能够原代培养大鼠远端PVSMC,所获细胞数量多、纯度高.

Culture of rat distal pulmonary venous smooth muscle cells by enzymatic digestion

Objective：To establish a method to culture pulmonary venous smooth muscle cells （PVSMC） from rat distal pulmonary veins for in vitro researches of the pulmonary circulation diseases. Methods： Microsurgery operations and collagenase I digestion were performed to isolate and culture rat distal PVSMC. The rat distal PVSMC were identified by morphological activity and immunofluorescence of smooth muscle α-actin. The purity of the ceils was checked by laser-scanning confocal fluorescence microscope. Results：The rat distal PVSMC displayed typically ＂peak and valley＂ growth pattern and expressed the smooth muscle specific differentiation marker smooth muscle α-actin. The purity of PVMC reached 98.5%. Conclusion： The rat distal PVSMC can be primarily cultured. PVSMC were bigger in quantity and high purity by this method.